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Two Hemicycliophora species, H. lutosa and H. typica, found in samples from fallow soil and sugarcane soil in South Africa, were studied morphologically and, for the first time, molecularly. Diagnostic PCR-IT-rRNA-RFLP profiles generated by five restriction enzymes are provided. Study of phylogenetic relationships using D2-D3 expansion segment of 28S rRNA gene sequences revealed that H. lutosa was related to H. poranga. Hemicycliophora lutosa and H. poranga are compared morphologically. SEM photographs are given for H. typica and for H. lutosa for the first time. The male of H. typica represents a first report for South Africa.